SUMMARY NAD depletion occurs after excitotoxic insults and oxidative stress and causes neuronal death in several rodent models of neurodegenerative conditions, including models for brain ischemia/reperfusion injury and Wallerian degeneration. We have recently discovered that NAD depletion is also the primary cause of neuronal death induced by exposure to a misfolded and toxic form of the amyloidogenic prion protein (TPrP). NAD replenishment reversed the fate of TPrP-injured neurons in culture and improved motor function in a mouse model of prion disease. Therefore, our study established NAD restoration as a new therapeutic target for protein misfolding neurodegenerative diseases, a family of diseases that includes Alzheimer's, Parkinson's, prion diseases and amyotrophic lateral sclerosis (ALS). We developed a high- throughput screening strategy to identify NAD-dependent neuroprotective small molecules in the misfolded protein toxicity paradigm and then conducted a screening campaign. We identified and selected four chemical series providing complete neuroprotection and preserving NAD levels with nanomolar potency. One compound was tested in a murine model of ALS and improved the mice's muscle strength and motor function. We now aim to optimize these compounds for potency, selectivity towards NAD restoration and absence of undesired off-target effects, as well as favorable drug metabolism and pharmacokinetic properties including brain penetration. We will perform iterative rounds of structure-activity relationship studies and subject analogs from each scaffold to a series of in vitro and in vivo assays for prioritization within and between each lead series. This workplan will deliver novel small molecule neuroprotectants poised for late development and investigational new drug (IND)?enabling studies and ultimately for the treatment of severely debilitating neurological conditions linked to a deficit in NAD.